Method for establishing recurrent cervical cancer PDX model

Abstract

The invention discloses a method for establishing a recurrent cervical cancer PDX model, which relates to the technical field of biomedicine, and comprises the following three models: a cervical cancer PDX subcutaneous transplantation tumor model, a cervical cancer PDX orthotopic transplantation tumor model and a cervical cancer PDX recurrent metastatic tumor model. The cervical cancer PDX model is derived from an uncultured human tumor, and the consistency of the PDX model and a primary tumor sample is better shown under the condition of retaining histopathology, specific protein expression and molecular characteristics. The method can be used for truly reflecting the clinical medication effect, constructing a cervical cancer PDX model platform and developing individualized treatment and novel treatment strategies of cervical cancer patients according to the platform, has an important guiding value for solving the difficult-to-treat problem of advanced cervical cancer, and has a good application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of tumor research and treatment, and in particular to a method for establishing a recurrent cervical cancer PDX model. Background Art

[0002] Cervical cancer is one of the common malignant tumors of the female reproductive system. Its incidence rate ranks fourth among female malignant tumors. More than 310,000 people die from this disease each year. The 5-year survival rate of patients with recurrent or metastatic cervical cancer is only 17%, and the prognosis is poor. For recurrent or metastatic advanced cervical cancer, traditional treatments are surgery and radiotherapy and chemotherapy. However, due to the limitations of traditional treatments, the treatment efficacy of advanced cervical cancer is still poor. In order to improve the treatment efficacy of patients with advanced cervical cancer, people are gradually paying attention to the precise treatment plan based on the heterogeneity of the patient's tumor.

[0003] Finding a suitable preclinical model to predict patient treatment outcomes and conduct precision treatment is crucial to improving the treatment effect of advanced cervical cancer. According to previous studies, the commonly used model for evaluating drug efficacy is the cell line. After multiple passages in vitro, the important genetic information and biological characteristics of the tumor cell line have changed from the original tumor tissue, leading to doubts about its value in guiding clinical treatment. In contrast, the tumor cells of the PDX model come from clinical patients, which can more accurately retain the genetic complexity and biological characteristics of the patient's tumor and maintain the heterogeneity of the tumor. By establishing the association between the treatment phenotype and the drug resistance mechanism, the PDX model can help patients with advanced tumors to conduct precise drug guidance, predict the adaptive targets of the drug, and ultimately be used for drug safety and efficacy evaluation.

[0004] The PDX model is established by implanting patient-derived tumor tissue into severely immunodeficient mice. The main advantage of this type of model is that it can retain the characteristics of the patient's tumor tissue to a large extent; even after multiple passages, it can still maintain consistency with the patient's primary tumor tissue. Compared with currently commonly used preclinical models, such as organoids, PDX models can more effectively reflect the pharmacokinetic process, toxicity and side effects of drugs in the body, as well as their interactions with the tumor microenvironment.

[0005] Therefore, constructing a cervical cancer PDX model platform and developing personalized treatment and new treatment strategies for cervical cancer patients based on this platform is crucial to solving the problem of intractable disease in advanced cervical cancer. Summary of the invention

[0006] The purpose of the present invention is to address the deficiencies in the existing technology and provide a method for establishing a recurrent cervical cancer PDX model, thereby stably constructing a recurrent cervical cancer PDX model to lay the foundation for screening more effective anti-cervical cancer therapeutic drugs and formulating personalized treatment plans.

[0007] To achieve the above object, the technical solution of the present invention is to provide a method for establishing a recurrent cervical cancer PDX model, comprising the following steps:

[0008] Extract primary cervical cancer tissue from patients;

[0009] Immunodeficient mice were taken and inoculated with primary tissues;

[0010] Tumor volume reaches 100mm within 6 months of screening 3 The mice with tumors were designated as P1 generation. When the tumor volume grew to 500-1000mm 3 When the tumor volume grows to 500-1000mm, the cells are transplanted into new immunodeficient mice as P2 generation. 3 When the tumor was 40 μg / cm3, the mice were killed, the tumor was removed, and the tumor was divided into 4 parts, one of which was transplanted into the next immunodeficient mouse. The operation was repeated until the P4 generation, and a recurrent cervical cancer PDX model was obtained.

[0011] Wherein, the inoculated primary tissue is subjected to any of the following operations:

[0012] The primary tissue was subcutaneously inoculated in the two ribs to establish a cervical cancer PDX subcutaneous transplant tumor model;

[0013] The primary tissue is inoculated into the uterus to establish an orthotopic transplant tumor model of cervical cancer PDX;

[0014] The primary tissue was inoculated subcutaneously in the ribs and the tumor was grown to 500-1000 mm. 3 The cervical cancer PDX recurrent metastatic tumor model was established after the recurrence of the original tumor site.

[0015] Preferably, the remaining three samples of each generation are subjected to the following operations: RNA and DNA extraction; fixation with 4% paraformaldehyde and then embedding in paraffin; cutting into 2×2×2 mm tissue blocks and storing in liquid nitrogen using cell freezing solution.

[0016] Furthermore, the primary tissue is subcutaneously inoculated in the two ribs, specifically: the mouse is anesthetized by isoflurane inhalation, the mouse is placed in a lateral position, the two ribs are disinfected, the primary tissue that has been washed and cut into small pieces is subcutaneously inoculated in the two ribs, and the wound is disinfected.

[0017] Furthermore, the uterus is inoculated with the primary tissue, specifically as follows: mice are anesthetized intraperitoneally with 60 mg / kg of sodium pentobarbital, placed in a supine position, and the hair on the lower abdomen is shaved and disinfected; a longitudinal incision is made in the middle of the mouse's lower abdomen, and the skin is pulled to both sides, and the peritoneum is lifted with ophthalmic non-destructive forceps and cut open to enter the abdominal cavity, and the mouse Y-shaped uterus is initially found behind the bladder, and the uterus is carefully pulled out outside the peritoneum with microscopic non-destructive forceps, and the primary tissue is sutured to the lower end of the Y-shape; the uterus is returned to the abdominal cavity, the peritoneum and skin are sutured, and the skin is disinfected.

[0018] Furthermore, the primary tissue is subcutaneously inoculated in the two ribs, and when the tumor grows to 500-1000 mm 3 After that, when the recurrent tumor grows in the original tumor site, the specific steps are as follows: the primary tissue is washed and placed in a 24-well plate and cut into small pieces for later use; the mouse is anesthetized by isoflurane inhalation, the mouse is placed in a lateral position, the ribs are disinfected, and the washed and cut small pieces of primary tissue are inoculated subcutaneously in the ribs, and the wound is disinfected; when the tumor grows to 500-1000mm 3 At 4 pm, mice were anesthetized with 60 mg / kg sodium pentobarbital intraperitoneally, placed in a lateral decubitus position, and the hair on both sides of the ribs was shaved and disinfected. The sides of the mice were cut open, and the skin was pulled to both sides to expose the tumor tissue. The tumor was completely separated with blunt force, and the skin was sutured and disinfected. The mice were left to wait for a recurrent tumor to grow at the original tumor site before proceeding to the next step.

[0019] Preferably, the process of subculturing further comprises: when the tumor volume reaches 1000-1500 mm 3 Or when there is an ulcer on the surface, the tumor is surgically removed, a part is taken for case detection and gene sequencing, a part is frozen in cryopreservation protection solution, and a part is continued to be passaged.

[0020] Preferably, the immunodeficient mice are NOD-SCID mice, 5 to 8 weeks old.

[0021] The present invention also provides a human cervical cancer tissue standardized model library, which is prepared by the following steps:

[0022] The human cervical cancer tissue was cryopreserved as the first part;

[0023] Fixing and embedding human cervical cancer tissue as the second part;

[0024] As the third part, human cervical cancer tissues were inoculated into immunodeficient mice;

[0025] The PDX model established by the above-mentioned method for establishing a recurrent cervical cancer PDX model is cryopreserved and passaged as the fourth part;

[0026] The first part was subjected to gene molecular genetics identification; the second part was subjected to pathology and immunohistochemical staining identification; the third part was subjected to pathology, immunohistochemical staining, gene molecular genetics, gene mutation and heterogeneity identification; based on the identification results, the three parts were determined to be used to construct a standardized model library of xenotransplanted human cervical cancer.

[0027] The present invention also provides an application of the above-mentioned human cervical cancer tissue standardized model library in cervical cancer pathology research, drug use and treatment.

[0028] The mice were observed twice a week, the tissue texture was touched by hand, the tumor size was measured with a vernier caliper, and the tumor growth curve was recorded.

[0029] The beneficial effects of the present invention are:

[0030] The present invention overcomes the shortcomings of traditional cervical cancer animal models by establishing a recurrent cervical cancer PDX model, and constructs a recurrent cervical cancer PDX model with stable passage. This model can be used to simulate the growth of tumors in the human body, and has important guiding value for the occurrence and development mechanism of tumors, the screening and evaluation of anti-cervical cancer drugs, and the individualized formulation of treatment plans, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 The present invention provides a flow chart for establishing a subcutaneous tumor model of cervical cancer; wherein: (1) obtaining primary tissue from the patient; (2) cutting the primary tumor tissue from the patient into pieces; (3) subcutaneously transplanting it into NOD / SCID mice; (4) observing tumor growth; (5) dissecting tumor tissue blocks; (6) cutting the tumor tissue blocks of the P1 generation PDX model into small pieces; (7) repeating steps 3 to 6 and passaged to subsequent mice; and (8) storing the tissue blocks in liquid nitrogen.

[0032] Figure 2 This is the in situ model of cervical cancer of the present invention; wherein, (A) is the in situ model production process; and (B) is PET-CT tracing of the in situ tumor position.

[0033] Figure 3 This is the cervical cancer recurrence model of the present invention;

[0034] Figure 4 is the tumor formation time of the cervical cancer PDX model F1 to F4 generations of the present invention;

[0035] Figure 5HE staining and key immune index identification diagram of the cervical cancer PDX model of the present invention; wherein, (A) is a comparison of the pathological histology and protein marker consistency of the patient and the corresponding PDX model (generation P1 to P4); (B) is a schematic diagram of the division of epithelial and stromal components of PT and corresponding PDX P1 tumors, the green part represents the epithelial component, and the red part represents the stromal component; (C) is a statistical diagram of epithelial and stromal components of PT and corresponding P1 generation tumors (scale bar, 50 μm);

[0036] Figure 6 The whole exome sequencing analysis results of the cervical cancer PDX model of the present invention are shown, and the somatic mutations of cervical cancer-related genes are shown; wherein, (A) is the CNA amplification and deletion of the genome of the patient and the matched PDX model, (B) is the copy number difference between the corresponding patient PT of CAT082 and its corresponding PDX model P1 and P2, (C) is the copy number difference between CAT061 PDX P1 to P4, (D) is the statistical inconsistency of the copy number variation between the patient PT and P1, and P1 and P2, (E) is the comparison of the SNP consistency between the patient tumor and the corresponding PDX model, (F) is the comparison of the selected cancer-related genes and tumor mutation load consistency between the patient tumor and the corresponding PDX model, and (G) is the change of AID / APOBEC mutation features (features 2 and 13) in CAT082 and CAT145 patient tumors. DETAILED DESCRIPTION

[0037] The content of the present invention is further described below in conjunction with specific implementation cases and drawings, but the present invention is not limited to the following implementation cases. Among them, the recurrent cervical cancer PDX model proposed in the present invention includes a cervical cancer PDX subcutaneous transplant tumor model, a cervical cancer PDX orthotopic transplant tumor model, and a cervical cancer PDX recurrent metastatic tumor model.

[0038] Experimental animals: NOD-SCID mice, female, 5-8 weeks old, weighing 16-22 g, were housed in an SPF (specific pathogen-free) experimental animal barrier system.

[0039] Example 1

[0040] Establishing a cervical cancer PDX subcutaneous transplant tumor model includes the following steps (such as Figure 1 shown):

[0041] S1: Acquisition, preservation, transportation and processing of primary cervical cancer tissues: During the surgery of cervical cancer patients, part of the tumor tissue and adjacent tissues are removed, placed in a sterile 15ml centrifuge tube, the tissue is completely covered with tissue preservation solution, and placed in a thermos bucket filled with ice packs. Within 2 to 6 hours after the tissue is removed from the body, the tissue block is rinsed with PBS (containing 1% penicillin and streptomycin) in a clean bench to remove the vascular connective tissue on the tissue. The collected specimens are uniformly divided into 3 parts for processing. One part is about 5×5×5mm for RNA and DNA extraction; one part is about 3×3×3mm for 4% paraformaldehyde fixation and then used for paraffin tissue sectioning for histological analysis; one part is 3×3×3mm for PDX model establishment;

[0042] S2: Transplantation of primary tissue: Wash the primary tissue and place it in a 6-well plate, cut it into small pieces for later use; anesthetize the mice by inhalation of isoflurane, place them in a lateral position, disinfect the two ribs with conventional iodine and 75% alcohol, put the tumor tissue block into the cannula needle and implant it subcutaneously, inoculate it subcutaneously in the two ribs, disinfect the wound again with iodine, wait for the mice to wake up naturally, name them P1 mice, and closely observe the tumor growth of the mice after surgery;

[0043] S3: PDX tumor generation, cryopreservation and identification: The primary tumor tissue of the patient is PT generation, transplanted into mice, and within 6 months, the tumor volume reaches 100mm 3 , defined as tumor formation, this generation model is P1 generation, when the tumor volume grows to 500-1000mm 3 The tumor was transplanted into new immunodeficient mice, named P2 generation. The tumor-bearing mice were killed by isoflurane gas anesthesia. The skin around the tumor of the mouse was disinfected in the clean bench, and the mouse was dissected to remove the tumor. The tumor was divided into 4 parts, one of which was transplanted into the next mouse; one was used for RNA and DNA extraction; one was fixed with 4% paraformaldehyde for paraffin embedding; one was cut into several 2×2×2mm tissue blocks and stored in liquid nitrogen with cell freezing solution for subsequent experimental recovery. By analogy, P2, P3 and P4 generation cervical cancer mouse models were established. At the same time, the P2-P4 generation mouse models were divided into 4 parts for subsequent identification. No more transplantation was performed after transplantation to P4 generation.

[0044] Example 2

[0045] Establishment of cervical cancer PDX orthotopic xenograft tumor model includes the following steps:

[0046] S1: Treat tumor tissues in the same manner as described above for establishing a cervical cancer PDX subcutaneous transplant tumor model;

[0047] S2: Mice were anesthetized with 60 mg / kg sodium pentobarbital intraperitoneally and placed in a supine position. The hair on the lower abdomen was removed, disinfected with iodine, and then deiodinated with alcohol. A longitudinal incision of about 1 cm was made in the middle of the lower abdomen of the NOD-SCID mouse. The skin was pulled to both sides to expose the lower peritoneum. The peritoneum was gently lifted with non-destructive ophthalmic forceps and cut into the abdominal cavity. The Y-shaped uterus of the mouse was initially found behind the bladder. The uterus was carefully pulled out of the peritoneum with micro-non-destructive forceps, and the tissue block was sutured to the lower end of the Y-shape with 6-0 thread. The uterus was returned to the abdominal cavity, and the peritoneum and skin were sutured with absorbable surgical sutures. The skin was disinfected with iodine and 75% alcohol. The mice were allowed to wake up naturally and the growth of the mice after surgery was closely observed (e.g. Figure 2 A);

[0048] S3: The same method as above for the passage and cryopreservation of the cervical cancer PDX subcutaneous transplant tumor model was used to treat the P1 generation tumor tissue;

[0049] S4: Validation of the in situ model: feasible PET-CT ( Figure 2 B) and pathological verification of the in situ tumor tissue after in situ dissection.

[0050] Example 3

[0051] Establishing a cervical cancer PDX recurrent xenograft model includes the following steps (such as Figure 3 shown):

[0052] S1: Treat tumor tissues in the same manner as described above for establishing a cervical cancer PDX subcutaneous transplant tumor model;

[0053] S2: Transplantation of recurrent tissue: Wash the primary tissue and place it in a 6-well plate, cut it into small pieces for later use; anesthetize the mouse by isoflurane inhalation, place it in a lateral position, disinfect the ribs with conventional iodine and 75% alcohol, put the tumor tissue block into the cannula needle and implant it subcutaneously, inoculate it subcutaneously in the ribs, disinfect the wound again with iodine, wait for the mouse to wake up naturally, name it P1 generation mouse, and closely observe the tumor growth of the mouse after surgery; when the tumor grows to 500-1000mm 3 At 14:00, mice were anesthetized with 60 mg / kg sodium pentobarbital intraperitoneally, placed in a lateral position, and the hair on both sides of the ribs was removed. After disinfection with iodine, the skin was deiodinated with alcohol. A 1 cm long cut was made on the side of the NOD-SCID mouse, and the skin was pulled to both sides to expose the tumor tissue. The tumor was completely separated with blunt force, and the skin was sutured with surgical sutures. The skin was disinfected with iodine and 75% alcohol. The mice were allowed to wake up naturally and their growth was closely observed after surgery. After 1-3 months, recurrent tumors grew at the original tumor site of the mice, and subsequent experimental operations were feasible.

[0054] S3: The P1 generation tumor tissue was processed using the same method as S1 for the subcutaneous transplantation model of cervical cancer PDX as described above.

[0055] The present invention also provides a human cervical cancer tissue standardized model library, which is prepared by the following steps:

[0056] The human cervical cancer tissue was cryopreserved as the first part;

[0057] Fixing and embedding human cervical cancer tissue as the second part;

[0058] As the third part, human cervical cancer tissues were inoculated into immunodeficient mice;

[0059] The recurrent cervical cancer PDX model was cryopreserved and passaged as the fourth part.

[0060] In addition, it also includes the identification of the standardized model library of human cervical cancer tissue, specifically: genetic molecular genetics identification of the first part; pathological and immunohistochemical staining identification of the second part; pathological, immunohistochemical staining, genetic molecular genetics, gene mutation and heterogeneity identification of the third part; based on the identification results, the three parts are determined to be used to construct a standardized model library of human cervical cancer for xenotransplantation.

[0061] The three models were characterized by xenograft tumor models, including the following steps:

[0062] S1: The primary tumor tissue of the patient is taken as the PT generation, and transplanted into the mouse model as the P1 generation, and so on, until the transplantation to the P4 generation.

[0063] S2: Pathological verification: H&E staining and key proteochemical staining were performed to detect whether the histopathological characteristics of the tumor were consistent with those of the primary tumor tissue ( Figure 5 A), detected the proportion of cancer and stroma in tumor tissue, and observed that when the cells were passaged into mice, the tumor proportion decreased and the stroma increased ( Figure 5 BC);

[0064] S3: Molecular feature validation: GISTIC algorithm was used to analyze copy number changes and compare amplification and deletion of 10 matched PT and PDX P1 models. The results showed that PDX P1 and PT have similar characteristics ( Figure 6 In addition, this patent also uses the CN ratio algorithm to quantify copy number variation. The results show that the heterogeneity of PT and P1 in CNA results is 18.79% (3.03%-32.8%), the heterogeneity of P1 and P2 is 3.21% (1.65%-4.77%), and the heterogeneity of P1 and P4 is 2.1% ( Figure 6 BD). This patent further analyzed the somatic mutation spectrum of early passage (P1) and late passage (P4). The results of single cell nucleotide variation (SNP) suggest that PT and P1 maintain a high degree of consistency ( Figure 6 EG).

[0065] The standardized model library of source cervical cancer tissue described in the present invention can be widely used in cervical cancer pathology research, drug use and treatment: the model library can more accurately simulate the biological behavior and pathological changes of cervical cancer in the human body, providing researchers with an experimental platform close to the real situation; as a "living" platform for drug screening, it can more quickly evaluate the inhibitory effect and mechanism of action of candidate drugs on cervical cancer cells; research based on the model library can predict the response and prognosis of different patients to specific treatment plans, providing a scientific basis for doctors to formulate personalized treatment plans.

[0066] The above embodiments are only used to illustrate the present invention, but are not used to limit the scope of implementation of the present invention.

Claims

1. A method for establishing a recurrent cervical cancer PDX model, characterized in that: The following steps are involved: Extract primary cervical cancer tissue from patients; Immunodeficient mice were taken and inoculated with primary tissues; Tumor volume reaches 100mm within 6 months of screening 3 The mice were designated as P1 generation; when the tumor volume grew to 500-1000mm 3 When the tumor volume grows to 500-1000mm, the cells are transplanted into new immunodeficient mice as P2 generation. 3 When the tumor was 40 μg / cm3, the mice were killed, the tumor was removed, and the tumor was divided into 4 parts, one of which was transplanted into the next immunodeficient mouse. The operation was repeated until the P4 generation, and a recurrent cervical cancer PDX model was obtained. Wherein, the inoculated primary tissue is subjected to any of the following operations: The primary tissue was subcutaneously inoculated in the two ribs to establish a cervical cancer PDX subcutaneous transplant tumor model; The primary tissue is inoculated into the uterus to establish an orthotopic transplant tumor model of cervical cancer PDX; The primary tissue was inoculated subcutaneously in the ribs and the tumor was grown to 500-1000 mm. 3 The cervical cancer PDX recurrent metastatic tumor model was established after the recurrence of the original tumor site.

2. The method for establishing a recurrent cervical cancer PDX model according to claim 1, characterized in that: The remaining three samples of each generation were subjected to the following operations: RNA and DNA extraction; fixation with 4% paraformaldehyde and then embedding in paraffin; cutting into 2×2×2mm tissue blocks and storing in liquid nitrogen using cell freezing solution.

3. The method for establishing a recurrent cervical cancer PDX model according to claim 1, characterized in that: The subcutaneous inoculation of the primary tissue in the two ribs is specifically as follows: the mouse is anesthetized by inhalation of isoflurane, the mouse is placed in a lateral position, the two ribs are disinfected, the primary tissue that has been washed and cut into small pieces is subcutaneously inoculated in the two ribs, and the wound is disinfected.

4. The method for establishing a recurrent cervical cancer PDX model according to claim 1, characterized in that: The primary tissue is inoculated into the uterus, specifically: mice are anesthetized with 60 mg / kg sodium pentobarbital intraperitoneally, placed in a supine position, and the hair on the lower abdomen is removed and disinfected; the mouse's lower abdomen is cut longitudinally in the middle, the skin is pulled to both sides, the peritoneum is lifted with ophthalmic non-destructive forceps and cut into the abdominal cavity, the mouse Y-shaped uterus is initially found behind the bladder, the uterus is carefully pulled out of the peritoneum with micro-non-destructive forceps, and the primary tissue is sutured to the lower end of the Y-shape; the uterus is returned to the abdominal cavity, the peritoneum and skin are sutured, and the skin is disinfected.

5. The method for establishing a recurrent cervical cancer PDX model according to claim 1, characterized in that: The primary tissue was inoculated subcutaneously in the two ribs, and the tumor was grown to 500-1000 mm. 3 After that, when the recurrent tumor grows in the original tumor site, the specific steps are as follows: the primary tissue is washed and placed in a 24-well plate and cut into small pieces for later use; the mouse is anesthetized by isoflurane inhalation, the mouse is placed in a lateral position, the ribs are disinfected, and the washed and cut small pieces of primary tissue are inoculated subcutaneously in the ribs, and the wound is disinfected; when the tumor grows to 500-1000mm 3 At 4 pm, mice were anesthetized with 60 mg / kg sodium pentobarbital intraperitoneally, placed in a lateral decubitus position, and the hair on both sides of the ribs was shaved and disinfected. The sides of the mice were cut open, and the skin was pulled to both sides to expose the tumor tissue. The tumor was completely separated with blunt force, and the skin was sutured and disinfected. The mice were left to wait for a recurrent tumor to grow at the original tumor site before proceeding to the next step.

6. The method for establishing a recurrent cervical cancer PDX model according to claim 1, characterized in that: The process of subculturing also includes: when the tumor volume reaches 1000-1500mm 3 Or when there is an ulcer on the surface, the tumor is surgically removed, a part is taken for case detection and gene sequencing, a part is frozen in cryopreservation protection solution, and a part is continued to be passaged.

7. The method for establishing a recurrent cervical cancer PDX model according to claim 1, characterized in that: The immunodeficient mice are NOD-SCID mice, with an age of 5 to 8 weeks.

8. A standardized model library of human cervical cancer tissue, characterized in that: Prepared by the following steps: The human cervical cancer tissue was cryopreserved as the first part; Fixing and embedding human cervical cancer tissue as the second part; As the third part, human cervical cancer tissues were inoculated into immunodeficient mice; The PDX model established by the method for establishing a recurrent cervical cancer PDX model according to claim 1 is cryopreserved and passaged as the fourth part; The first part was subjected to gene molecular genetics identification; the second part was subjected to pathology and immunohistochemical staining identification; the third part was subjected to pathology, immunohistochemical staining, gene molecular genetics, gene mutation and heterogeneity identification; based on the identification results, the three parts were determined to be used to construct a standardized model library of xenotransplanted human cervical cancer.

9. Use of the standardized model library of human cervical cancer tissue according to claim 8 in cervical cancer pathology research, drug use and treatment.

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